Erratum: Author affiliation addition: "Hepatitis B virus detected in paper currencies in a densely populated city of India: A plausible source of horizontal transmission?"

[This corrects the article on p. 775 in vol. 12, PMID: 33200016.].

Microbial surfactants of marine origin: potentials and prospects.

Marine environment occupies the vast majority of the earth's surface and is a rich source of highly potent and active compounds. In recent years, microbial surfactants and emulsifiers have been reported from marine microflora. Surfactant and emulsifier molecules having diverse chemical nature such as exopolysaccharides, carbohydrate-lipid-protein complexes or glycolipopeptide, glycolipids, lipopeptides, phospholipids and ornithine lipids have been reported from various marine bacteria. These surface-active agents have been found to possess good emulsification and stabilization potentials for various lipophilic compounds such as aliphatic, aromatic and polyaromatic hydrocarbons and their uptake and degradation by the microorganisms. Few biosurfactant types such as glycolipids and lipopeptides have also been found to possess valuable biological activities. Surface-active agents from marine environments thus have tremendous potential to be used in industrial processes, for environmental remediation and as drugs.

Hepatitis B virus detected in paper currencies in a densely populated city of India: A plausible source of horizontal transmission?

BACKGROUND: The recent rise in the incidence of hepatitis B virus (HBV) infections in a densely populated city of eastern India ("mixing vessel" of people of varied socio-economic and immune status) prompted this study. Applying saliva on fingers for enumerating bank notes is a common practice in the Indian subcontinent. Paper notes may be a potential source of "horizontal" transmission of this virus, especially if there are cuts/bruises on the oral mucous membrane or skin. AIM: To investigate whether paper currencies could be a plausible mode of horizontal transmission of HBV infection. METHODS: Polymerase chain reactions (PCR) followed by nucleotide sequencing was done for the detection of HBV. Hepatitis B virus surface antigen enzyme-linked immunosorbent assay(HBsAg ELISA) was performed on all HBV deoxyribonucleic acid-positive samples to check the detectability of the virus. Atomic force microscopy (AFM) was carried out for visual confirmation of HBV particles in ultracentrifuged/immunoprecipitated samples from currency paper washings. RESULTS: HBV-specific PCRs on pellets obtained after ultracentrifugation/ immunoprecipitation of the currency paper washings detected potentially intact/viable HBV (genotype D2) in 7.14% of samples (n = 70). AFM gave the visual confirmation of HBV particles in ultracentrifuged/immunoprecipitated samples from currency paper washings. However, HBV isolates from the currency notes could not be detected by HBsAg ELISA. CONCLUSION: It is a common practice in the Indian subcontinent to count paper currencies by applying saliva on fingertips. Paper notes may be a potential source of "horizontal" transmission of this virus, especially if there are cuts/bruises on the oral mucous membrane or skin, but it was practically not possible to demonstrate experimentally such transmission. Detection of potentially intact/viable and "occult" HBV from currency poses potential risk of silent transmission of this virus among the general population.

Rapid quantification of a microbial surfactant by a simple turbidometric method.

Quantification of the biosurfactants produced by a variety of microorganisms is a time taking and difficult task due to the lack of rapid, efficient and accurate methods. This work presents a simple turbidometric method for quantification of crude biosurfactants based on their property to become insoluble at low pH values. Biosurfactants obtained from a Bacillus sp. using different carbon substrates showed a good linear correlation (R(2)>0.99) between biosurfactant concentrations and turbidity in the range of 1 to 10 g L(-1) of crude biosurfactants. The substrate specific equations (SSE) and generalized equations (GE) developed in this work effectively predicted the amount of crude biosurfactant produced in different sets of fermentation experiments validating the method. A similar linear correlation was also observed with biosurfactants obtained from two other strains, Bacillus circulans and Pseudomonas sp. This simple method may prove to be effective in fast, accurate and inexpensive quantification of crude biosurfactants produced by diverse bacteria.

New avenues of controlling microbial infections through anti-microbial and anti-biofilm potentials of green mono-and multi-metallic nanoparticles: A review.

Nanoparticles synthesized through the green route deserve special mention because this green technology is not only energy-efficient and cost-effective but also amenable to the environment. Various biological resources have been used for the generation of these 'green nanoparticles'. Biological wastes have also been focused in this direction thereby promoting the value of waste. Reports indicate that green nanoparticles exhibit remarkable antimicrobial activitiesboth singly as well as in combination with standard antibiotics. The current phenomenon of multi-drug resistance has resulted due to indiscriminate administration of high-doses of antibiotics followed by significant toxicity. In the face of this emergence of drug-resistant microbesthe efficacy of green nanoparticles might prove greatly beneficial. Microbial biofilm is another hurdle in the effective treatment of diseases as the microorganismsbeing embedded in the meshwork of the biofilmevade the antimicrobial agents. Nanoparticles may act as a ray of hope on the face of this challenge tooas they not only destroy the biofilms but also lessen the doses of antibiotics requiredwhen administered in combination with the nanoparticles. It should be further noted that the resistance mechanisms exhibited by the microorganisms seem not that relevant for nanoparticles. The current review, to the best of our knowledgefocuses on the structures of these green nanoparticles along with their biomedical potentials. It is interesting to note how a variety of structures are generated by using resources like microbes or plants or plant products and how the structure affects their activities. This study might pave the way for further development in this arena and future work may be taken up in identifying the detailed mechanism by which 'green' synthesis empowers nanoparticles to kill pathogenic microbes.

Improved bioavailability and biodegradation of a model polyaromatic hydrocarbon by a biosurfactant producing bacterium of marine origin.

Polyaromatic hydrocarbons (PAHs) are organic pollutants mostly derived from the processing and combustion of fossil fuels and cause human health hazards. In the present study a marine biosurfactant producing strain of Bacillus circulans was used to increase the bioavailability and consequent degradation of a model polyaromatic hydrocarbon, anthracene. Although the organism could not utilize anthracene as the sole carbon source, it showed better growth and biosurfactant production in an anthracene supplemented glycerol mineral salts medium (AGlyMSM) compared to a normal glycerol mineral salts medium (GlyMSM). The biosurfactant product showed high degree of emulsification of various hydrocarbons. Analysis by gas chromatography (GC), high performance thin layer chromatography (HPTLC) and Fourier transform infrared spectroscopy (FTIR) showed that the biosurfactant could effectively entrap and solubilize PAH. Thin layer chromatographic analysis showed that anthracene was utilized as a carbon substrate for the production of biosurfactant. Thus organic pollutant anthracene was metabolized and converted to biosurfactants facilitating its own bioremediation.

Genetic regulations of the biosynthesis of microbial surfactants: an overview.

Microbial biosurfactants are surface active metabolites synthesized by microbes growing on a variety of substrates. In spite of having great potential for commercial, therapeutic and environmental applications, industrial level production has not been realized for their low yields and productivities. One vital factor determining their biosynthesis is the genetic makeup of the producer organisms. Studies on molecular genetics and biochemistry of the synthesis of several biosurfactants have revealed the operons, the enzymes and the metabolic pathways required for their extracellular production. Surfactin, a cyclic lipopeptide biosurfactant is a potent antimicrobial agent and is produced as a result of non-ribosomal biosynthesis catalyzed by a large multienzyme peptide synthetase complex called the surfactin synthetase. Pathways for the synthesis of other lipopeptides such as iturin, lichenysin and arthrofactin are also mediated by similar enzyme complexes. These non-ribosomal peptide synthetases (NRPSs) responsible for lipopeptide biosynthesis display a high degree of structural similarity among themselves even from distant microbial species. Plasmid-encoded- rhlA, B, R and I genes of rhl quorum sensing system are required for production of glycolipid biosurfactants by Pseudomonas species. Molecular genetics of biosynthesis of alasan and emulsan by Acinetobacter species and of the fungal biosurfactants such as mannosylerythritol lipids (MEL) and hydrophobins have been deciphered. However, limited genetic information is available about biosynthesis of other biosurfactants such as viscosin, amphisin and putisolvin produced by some strains of Pseudomonas species. Understanding of the genetic regulatory mechanisms would help to develop metabolically engineered hyper-producing strains with better product characteristics and acquired capability of utilizing cheap agro-industrial wastes as substrates. This article thus provides an overview of the role and importance of molecular genetics and gene regulation mechanisms behind the biosynthesis of various microbial surfactants of commercial importance.

Towards commercial production of microbial surfactants.

Biosurfactants or microbial surfactants are surface-active biomolecules that are produced by a variety of microorganisms. Biosurfactants have gained importance in the fields of enhanced oil recovery, environmental bioremediation, food processing and pharmaceuticals owing to their unique properties--higher biodegradability, lower toxicity, and effectiveness at extremes of temperature, pH and salinity. However, large-scale production of these molecules has not been realized because of low yields in production processes and high recovery and purification costs. This article describes some practical approaches that have been adopted to make the biosurfactant production process economically attractive: these include the use of cheaper raw materials, optimized and efficient bioprocesses and overproducing mutant and recombinant strains for obtaining maximum productivity. The application of these strategies in biosurfactant production processes, particularly those using hyper-producing recombinant strains in the optimally controlled environment of a bioreactor, might lead towards the successful commercial production of these valuable and versatile biomolecules in near future.

Analysis of biosurfactants from industrially viable Pseudomonas strain isolated from crude oil suggests how rhamnolipids congeners affect emulsification property and antimicrobial activity.

Rhamnolipid biosurfactants produced mainly by Pseudomonas sp. had been reported to possess a wide range of potential industrial application. These biosurfactants are produced as monorhamnolipid (MRL) and di-rhamnolipid (DRL) congeners. The present study deals with rhamnolipid biosurfactants produced by three bacterial isolates from crude oil. Biosurfactants produced by one of the strains (named as IMP67) was found to be very efficacious based on its critical micelle concentration value and hydrocarbon emulsification property. Strikingly, antimicrobial, and anti-biofilm potential of this biosurfactant were higher than biosurfactants produced by other two strains. Thin layer chromatography analysis and rhamnose quantification showed that the rhamnolipids of IMP67 had more MRL congeners than biosurfactants of the other two strains. Emulsification and antimicrobial actions were affected by manual change of MRL and DRL congener proportions. Increase of MRL proportion enhanced emulsification index and antimicrobial property to Gram negative bacteria. This result indicated that the ratio of MRL and DRL affected the emulsification potentials of rhamnolipids, and suggested that high emulsification potentials might enhance rhamnolipids to penetrate the cell wall of Gram negative bacteria. In line with this finding, rhamnolipids of IMP67 also reduced the MIC of some antibiotics against bacteria, suggesting their synergistic role with the antibiotics.

Biosurfactant of marine origin exhibiting heavy metal remediation properties.

The present study was aimed at elucidating the role of biosurfactant product isolated from a marine bacterium in removing heavy metals from heavy metal containing solutions. In this study, metal removal was biosurfactant-mediated. Efficiency of metal removal depended on the concentration of the metal as well as that of the biosurfactant. At a concentration 5x, the critical micelle concentration (CMC), almost complete removal of 100 ppm of lead and cadmium occurred. Atomic absorption spectroscopy (AAS) studies also showed metal removal at a concentration less than the CMC in contrast to earlier findings that only micelles are involved in metal removal. Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM) equipped with energy dispersive X-ray spectroscopy (EDS) further substantiated these findings.

Substrate dependent production of extracellular biosurfactant by a marine bacterium.

The potential of a marine microorganism to utilize different carbon substrates for the production of an extracellular biosurfactant was evaluated. Among the several carbon substrates tested for this purpose, production of the crude biosurfactant was found to be highest with glycerol (2.9+/-0.11 g L(-1)) followed by starch (2.5+/-0.11 g L(-1)), glucose (1.16+/-0.11 g L(-1)) and sucrose (0.94+/-0.07 g L(-1)). The crude biosurfactant obtained from glycerol, starch and sucrose media had significantly higher antimicrobial action than those obtained from glucose containing medium. RP-HPLC resolved the crude biosurfactants into several fractions one of which had significant antimicrobial action. The antimicrobial fraction was found in higher concentrations in biosurfactant obtained using glycerol, starch and sucrose as compared to the biosurfactants from glucose medium, thereby explaining higher antimicrobial activity. The carbon substrate was thus found to affect biosurfactant production both in a qualitative and quantitative manner.

Antiadhesive action of a marine microbial surfactant.

The antiadhesive action of a lipopeptide biosurfactant from a marine bacterium was investigated. The effect of cultivation conditions on the adhesion property of few bacterial strains was studied. It was observed that the static cultures showed greater adhesion due to scarcity of oxygen. The biosurfactant upon surface conditioning was found to be effective in removal of the microbial adhesion at a concentration as low as 0.1 g L(-1). The percentages of inhibition of adhesion against different test bacterial strains ranged from 15 to 89% using 0.1-10 g L(-1) of purified biosurfactant. These percentages of adhesion inhibition were found to be significantly higher than the previously reported values. The antiadhesive efficacy of the biosurfactant was also evident from confocal laser scanning microscopy studies.